A Visual Gene Array for the Detection of HSV-2 Based on Nanogold Probes

LIU Ming-gui; WANG Ye-fu; SUI Fang; WAN Zhi-xiang; GAO Wen-jun

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June 2006

LIU Ming-gui, WANG Ye-fu, SUI Fang, WAN Zhi-xiang and GAO Wen-jun. A Visual Gene Array for the Detection of HSV-2 Based on Nanogold Probes[J]. Virologica Sinica, 2006, 21(3): 294-297.

Citation: LIU Ming-gui, WANG Ye-fu, SUI Fang, WAN Zhi-xiang, GAO Wen-jun. A Visual Gene Array for the Detection of HSV-2 Based on Nanogold Probes .VIROLOGICA SINICA, 2006, 21(3) : 294-297.

应用纳米金探针检测HSV-2的目视化基因芯片

刘明贵 ¹ ,
王业富 ^1*,, ,
隋放 ² ,
万志香 ¹ ,
高文娟 ¹

1.
1.武汉大学生命科学学院病毒学国家重点实验室, 湖北武汉 430072
2.
辽阳市中心医院, 辽宁辽阳 111000

通讯作者： 王业富, ;

摘要

本实验建立了一种应用金标链霉亲和素探针的目视化高灵敏度检测单纯疱疹病毒2型（Herpes Simplex Virus-2, HSV-2）的基因芯片。该芯片以HSV-2 DNA 聚合酶的高保守区为靶序列，设计HSV-2特异性引物和探针，通过PCR反应使扩增产物标记上生物素；氨基修饰的探针固定在活化的玻片上，与生物素标记的扩增产物杂交；利用生物素与链酶亲和素高亲合力的特性，加入纳米金标记的链酶亲和素后形成生物素-链酶亲和素-纳米金生物反应放大系统；银染反应后，达到目视化检测HSV-2效果。该HSV-2检测基因芯片能目视化检测出100fmol/L的HSV-2扩增产物，具有灵敏度高，低成本的特点，通过临床标本验证表明该芯片具有较高的准确性。
- 基因芯片
- , HSV-2
- , 纳米金
- , 生物素-链霉亲和素
- , 灵敏度

A Visual Gene Array for the Detection of HSV-2 Based on Nanogold Probes

1.
1. State key Lab of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China
2.
Liaoyang Center Hospital, Liaoyang 111000, China

Corresponding author: WANG Ye-fu,

Abstract

A visual HSV-2 detection gene array was developed based on nanogold probes。The specific primer and probe of HSV-2 were designed based on the highly conservative region of HSV-2 DNA polymerase gene and the HSV-2 PCR amplicon were labeled by biotin through PCR reaction．Amino-modified probes were spotted on an activated glass surface on which hybridized with the PCR amplicon．Subsequently, the streptavidin-nanogold was added to form the amplification system of the biotin-streptavidin-nanogold complex．At last, the step of silver staining amplified the signal of hybridization, Therefore it was easy to detect the HSV-2 by naked eyes．As little as 100fmol/L of HSV-2 amplicon could be detected on the array, The HSV-2 detection gene array can detect the HSV-2 virus infection with high sensitivity and low cost.
- Gene array
- , HSV-2
- , Nanogold
- , Biotin-streptavidin
- , Sensitivity

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

A Visual Gene Array for the Detection of HSV-2 Based on Nanogold Probes

Corresponding author: WANG Ye-fu,

1. 1. State key Lab of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China
2. Liaoyang Center Hospital, Liaoyang 111000, China

Abstract: A visual HSV-2 detection gene array was developed based on nanogold probes。The specific primer and probe of HSV-2 were designed based on the highly conservative region of HSV-2 DNA polymerase gene and the HSV-2 PCR amplicon were labeled by biotin through PCR reaction．Amino-modified probes were spotted on an activated glass surface on which hybridized with the PCR amplicon．Subsequently, the streptavidin-nanogold was added to form the amplification system of the biotin-streptavidin-nanogold complex．At last, the step of silver staining amplified the signal of hybridization, Therefore it was easy to detect the HSV-2 by naked eyes．As little as 100fmol/L of HSV-2 amplicon could be detected on the array, The HSV-2 detection gene array can detect the HSV-2 virus infection with high sensitivity and low cost.